This application is filed pursuant to 35 U.S.C. xc2xa7371 as a United States National Phase Application of International Application No. PCT/EP00/05720 filed Jun. 22, 2000, which claims priority from GB9914977.5 filed Jun. 25, 1999 in the United Kingdom.
The present invention relates to certain novel compounds. In particular, the present invention relates to compounds that activate the delta subtype of the human peroxisome proliferator activated receptor (xe2x80x9chPPARxcex4xe2x80x9d). The present invention also relates to method for preparing and using the novel compounds and to methods for using activators of hPPARxcex4.
Several independent risk factors have been associated with cardiovascular disease. These include hypertension, increased fibrinogen levels, high levels of triglycerides, elevated LDL cholesterol, elevated total cholesterol, and low levels of HDL cholesterol. HMG CoA reductase inhibitors (xe2x80x9cstatinsxe2x80x9d) are useful for treating conditions characterized by high LDL-c levels. It has been shown that lowering LDL-c is not sufficient for reducing the risk of cardiovascular disease in some patients, particularly those with normal LDL-c levels. This population pool is identified by the independent risk factor of low HDL-c. The increased risk of cardiovascular disease associated with low HDL-c levels has not yet been successfully addressed by drug therapy (i.e. currently there are no drugs on the market that are useful for raising HDL-c). (Bisgaier, C. L.; Pape, M. E. Curr. Pharm. Des. 1998, 4, 53-70).
Syndrome X (including metabolic syndrome) is loosely defined as a collection of abnormalities including hyperinsulinemia, obesity, elevated levels of trigycerides, uric acid, fibrinogen, small dense LDL particles, and plasminogen activator inhibitor 1 (PAI-1), and decreased levels of HDL-c.
NIDDM (non insulin dependent or Type 2 diabetes mellitus) is described as insulin resistance which in turn causes anomalous glucose output and a decrease in glucose uptake by skeletal muscle. These factors eventually lead to impaired glucose tolerance (IGT) and hyperinsulinemia.
Three mammalian Peroxisome Proliferator-Activated Receptors have been isolated and termed PPAR-alpha, PPAR-gamma, and PPAR-delta (also known as NUC1 or PPAR-beta). These PPARs regulate expression of target genes by binding to DNA sequence elements, termed PPAR response elements (PPRE). To date, PPRE""s have been identified in the enhancers of a number of genes encoding proteins that regulate lipid metabolism suggesting that PPARs play a pivotal role in the adipogenic signaling cascade and lipid homeostasis (H. Keller and W. Wahli, Trends Endoodn. Met 291-296, 4 (1993)).
It has now been reported that thiazolidinediones are potent and selective activators of PPAR-gamma and bind directly to the PPAR-gamma receptor (J. M. Lehmann et. al., J. Biol. Chem. 12953-12956, 270 (1995)), providing evidence that PPAR-gamma is a possible target for the therapeutic actions of the thiazolidinediones.
Activators of the nuclear receptor PPARxcex3 gamma, for example troglitazone, have been shown in the clinic to enhance insulin-action, reduce serum glucose and have small but significant effects on reducing serum triglyceride levelsin patients with Type 2 diabetes. See, for example, D. E. Kelly et al., Curr. Opin. Endocrinol. Diabetes, 90-96, 5 (2), (1998); M. D. Johnson et al., Ann. Pharinacother., 337-348, 32. (3), (1997); and M. Leutenegger et al., Curr. Ther. Res., 403-416, 58 (7), (1997).
The mechanism for this triglyceride lowering effect appears to be predominantly increased clearance of very low density lipoproteins (VLDL) through induction of liporotein lipase (LPL) gene expression. See, for example, B. Staels et al., Aerieoscler. Thromb., Vasc. Biol., 1756-1764, 17 (9), (1997).
Fibrates are a class of drugs which may lower serum triglycerides 20-50%, lower LDLc 10-15%, shift the LDL particle size from the more atherogenic small dense to normal dense LDL, and increase HDLc 10-15%. Experimental evidence indicates that the effects of fibrates on serum lipids are mediated through activation of PPARxcex1. See, for example, B. Staels et al., Curr. Pharm. Des., 1-14, 3 (1), (1997). Activation of PPAR alpha results in transcription of enzymes that increase fatty acid catabolism and decrease de-novo fatty acid synthesis in the liver resulting in decreased triglyceride synthesis and VLDL production/secretion. In addition, PPAR alpha activation decreases production of apoC-III. Reduction in apoC-III, an inhibitor of LPL activity, increases clearance of VLDL. See, for example, J. Auwerx et al., Atherosclerosis, (Shannon, Irel.), S29-S37, 124 (Suppl), (1996).
Certain compounds that activate or otherwise interact with one or more of the PPARs have been implicated in the regulation of triglyceride and cholesterol levels in animal models. See, for example, U.S. Pat. No. 5,847,008 (Doebber et al.) and U.S. Pat. No. 5,859,051 (Adams et al.) and PCT publications WO 97/28149 (Leibowitz et al.) and WO 99/04815 (Shimokawa et al.). In a recent report (Berger et al., J. Biol. Chem. 1999), vol. 274, pp. 6718-6725) it was stated that PPAR delta activation does not appear to modulate glucose or triglyceride levels.
Briefly, in one aspect, the present invention. provides compounds of formula (I), and pharmaceutically acceptable salts and, solvates thereof, wherein 
X represents a COOH (or a hydrolysable ester thereof) or tetrazole group;
X1 represents NH, NCH3, O, S, a bond (i.e., is absent), CH2, or CH where the dashed line indicates that when X1 is CH the depicted bond is a double bond;
X2 represents O or S;
R1 and R2 independently represent H, CH3, OCH3, or halogen;
n is 1 or 2;
one of Y and Z is N and the other is S or O:
y is0 1, 2, 3, 4 or 5;
Each R3 independently represents CF3 or halogen.
In another aspect, the present invention discloses a method for prevention or treatment of a human PPAR delta (xe2x80x9chPPARxcex4xe2x80x9d) mediated disease or condition comprising administration of a therapeutically effective amount of a compound of this invention. hPPARxcex4 mediated diseases or conditions include dyslipidemia including associated diabetic dyslipidemia and mixed dyslipidemia, syndrome X (as defined in this application this embraces metabolic syndrome), heart failure, hypercholesteremia, cardiovascular disease including atherosclerosis, arteriosclerosis, and hypertriglyceridemia, Type 2 diabetes mellitus, Type 1 diabetes, insulin resistance, hyperlipidemia, and regulation of appetite and food intake in subjects suffering from disorders such as obesity, anorexia bulimia, and anorexia nervosa. In particular, the compounds of this invention are useful in the treatment and prevention of cardiovascular diseases and conditions including atherosclerosis, arteriosclerosis, hypertriglyceridemia, and mixed dyslipidaemia.
In another aspect, the present invention provides pharmaceutical compositions comprising a compound of the invention, preferably in association with a pharmaceutically acceptable diluent or carrier.
In another aspect, the present invention provides a compound of the invention for use in therapy, and in particular, in human medicine.
In another aspect, the present invention provides the use of a compound of the invention for the manufacture of a medicament for the treatment of a hPPARxcex4 mediated disease or condition.
In another aspect, the present invention provides a method for lowering triglycerides by administration of a hPPARxcex4 agonist. Preferably the hPPARxcex4 agonist is a selective agonist.
In another aspect the present invention provides the use of a hPPARxcex4 agonist for the manufacture of a medicament for lowering triglyceride. levels. Preferably the hPPARxcex4 agonist is a selective agonist.
In a further aspect the present invention provides a method for treating Type 2 diabetes, decreasing insulin resistance or lowering blood pressure comprising administering a hPPARxcex4 agonist. Preferably the hPPARxcex4 agonist is a selective agonist.
In a further aspect there is provided the use of a hPPARxcex4 agonist for the manufacture of a medicament for treating Type 2 diabetes, decreasing insulin resistance or lowering blood pressure. Preferably the hPPARxcex4 agonist is a selective agonist.
In a further aspect the invention provides a method for decreasing fibrinogen levels comprising administering a hPPARxcex4 agonist. Preferably the hPPARxcex4 agonist is a selective agonist.
In a further aspect there is provided the use of a hPPARxcex4 agonist for the manufacture of a medicament for decreasing fibrinogen levels. Preferably the hPPARxcex4 agonist is a selective agonist.
In a further aspect the invention provides a method for decreasing LDLc levels comprising administering a hPPARxcex4 agonist. Preferably the hPPARxcex4 agonist is a selective agonist.
In a further aspect the invention provides the use of a hPPARxcex4 agonist for the manufacture of a medicament for decreasing LDLc levels. Preferably the hPPARxcex4 agonist is a selective agonist.
In a further aspect the invention provides a method for shifting the LDL particle size from small dense to normal dense LDL comprising administering a hPPARxcex4 agonist. Preferably the hPPARxcex4 agonist is a selective agonist.
In a further aspect the invention provides the use of a hPPARxcex4 agonist for the manufacture of a medicament for shifting the LDL particle size from small dense to normal dense LDL. Preferably the hPPARxcex4 agonist is a selective agonist.
As used herein, xe2x80x9ca compound of the inventionxe2x80x9d means a compound of formula (I) or a pharmaceutically acceptable salt or, solvate, thereof.
While hydrolyzable esters and tetrazole derivatives are included in the scope of this invention, the acids are preferred because the data suggests that while the esters are useful compounds, it may actually be the acids to which they hydrolyze that are the active compounds. Esters that hydrolyze readily can produce the carboxylic acid in the assay conditions or in vivo. Generally the carboxylic acid is active in both the binding and transient transfection assays, while the ester does not usually bind well but is active in the transient transfection assay presumably due to hydrolysis. Preferred hydrolysable esters are C1-6, alkyl esters wherein the alkyl group may be straight chain or branched chain. Methyl or ethyl esters are more preferred.
Preferably therefore X represents COOH.
Preferably X1 is O, S, or is absent. More preferably X1 represents O.
Preferably X2 is S.
Preferably R1 is H or CH3, more preferably CH3.
Preferably R2 is H.
Preferably Z is N.
Preferably Y is S.
Preferably n is 1.
Preferably y is 1 or 2. When y is 2, preferably one of the substituents is halogen; more preferably one is halogen and the other is CF3. More preferably y is 1. When y is 1, preferably the substituent is in the para position on the ring and is more preferably CF3.
A particular group of compounds is compounds of formula (II), and pharmaceutically acceptable salts, solvates, and hydrolyzable esters thereof, wherein 
X1 is NH, NCH3, O, S, a bond (i.e. is absent), CH2, or CH where the dashed line indicates that when X1 is CH the depicted bond is a double bond;
X2 is O or S;
R1 is H, CH3, OCH3, or halogen;
R2 is H, OCH3, or halogen
n is 1 or 2;
one of Y and Z is N and the other is S or O.
R3 is H, CF3 or halogen.
While the preferred groups for each variable have generally been listed above separately for each variable, preferred compounds of this invention include those in which several or each variable in Formula (I) is selected from the preferred, more preferred, or most preferred groups for each variable. Therefore, this invention is intended to include all combinations of preferred, more preferred, and most preferred groups.
Preferably, the compounds of formula (I) are hPPARxcex4 agonists. As used herein, by xe2x80x9cagonistxe2x80x9d, or xe2x80x9cactivating compoundxe2x80x9d, or xe2x80x9cactivatorxe2x80x9d, or the like, is meant those compounds which have a pKi of at least 6.0, preferably at least 7.0, to the relevant PPAR, for example hPPARxcex4, in the binding assay described below, and which achieve at least 50% activation of the relevant PPAR relative to the appropriate indicated positive control in the transfection assay described below at concentrations of 10xe2x88x925 M or less. Preferably, the agonist of this invention achieve 50% activation of human PPARxcex4 in the transfection assay at concentrations of 10xe2x88x927 M or less, more preferably 10xe2x88x929M or less.
Most preferably, the compounds of formula (I) are selective hPPARxcex4 agonists. As used herein, a xe2x80x9cselective hPPARxcex4 agonistxe2x80x9d is a hPPARxcex4 agonist whose EC50 for PPARxcex4 is at least 10 fold lower than its EC50 for PPARxcex3 and PPARxcex1. Such selective compounds may be referred to as xe2x80x9c10-fold selective.xe2x80x9d EC50 is defined in the transfection assay described below and is the concentration at which a compound achieves 50% of its maximum activity. Most preferred compounds are greater than 100-fold selective hPPARxcex4 agonists.
The PPARxcex4 selective compounds of this invention elevate HDL-c in db/db mice and primate models and lower fibrinogen in primate models. These PPARxcex4 selective agonists unexpectedly lower triglycerides and insulin levels in the primate.
Since the literature suggests that such triglyceride and fibrinogen lowering effects are due to PPAR alpha agonist activity, it would not be obvious that adding PPAR delta agonist activity to other PPAR activity such as alpha or gamma or alpha/gamma dual activity, would provide any additional triglyceride or fibrinogen lowering benefits. We have surprisingly found that adding PPAR delta activity to other PPAR activity, including PPAR alpha activity, could result in additional triglyceride, LDLc; or fibrinogen lowering benefits as well as decreasing insulin resistance.
Preferred compounds of the present invention include:
2-[2-methyl-4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-oxazol-5-yl}methoxy)phenoxy]acetic acid
3-[2-methyl-4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methoxy)phenyl]propanoic acid
3-[2-methyl-4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-oxazol-5-yl}methoxy)phenyl]propanoic acid
2-{4-[({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-oxazol-5-yl}methyl)sulfanyl]phenyl}acetic acid
2-[2-methyl-4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methoxy)phenoxy]acetic acid
3-[4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methoxy)phenyl]propanoic acid
(E)-3-[2-methyl-4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-oxazol-5-yl}methoxy)phenyl]-2-propenoic acid
methyl 3-[4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methoxy)phenyl]propanoate
2-{4-[({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenyl}acetic acid
2-({4-[({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-oxazol-5-yl}methyl)sulfanyl]phenyl}sulfanyl)acetic acid
2-[methyl-4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methoxy)anilino]acetic acid
2-{3-chloro-4-[({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-oxazol-5-yl}methyl)sulfanyl]phenyl}acetic acid
2-[2-chloro-4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-oxazol-5-yl}methoxy)phenoxy]acetic acid
2-[3-chloro-4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methoxy)phenyl]acetic acid
2-{4-[({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-oxazol-5-yl}methyl)sulfanyl]phenoxy}acetic acid
(E)-3-[4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-oxazol-5-yl}methoxy)phenyl]-2-propenoic acid
2-[4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-oxazol-5-yl}methoxy)phenoxy]acetic acid
2-[3-fluoro-4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methoxy)phenyl]acetic acid
methyl 2-[3-chloro-4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-oxazol-5-yl}methoxy)phenyl]acetate
2-{2-methyl-4-[({4-methyl-2-[4-bromophenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetic acid
ethyl 2-{2-methyl-4-[({4-methyl-2-[4-bromophenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetate
2-{-2-methyl-4-[({4-methyl-2-[4-chlorophenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetic acid
ethyl 2-{2-methyl-4-[({4-methyl-2-[4-chlorophenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetate
2-{2-methyl-4-[({4-methyl-2-[4-fluorophenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetic acid
ethyl 2-{2-methyl-4-[({4-methyl-2-[4-fluorophenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetate
2-{2-methyl-4-[({4-methyl-2-[3,4-difluorophenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetic acid
ethyl 2-{2-methyl-4-[({4-methyl-2-[3,4-difluorophenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetate
2-{2-methyl-4-[({4-methyl-2-[3,4-dichlorophenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetic acid
ethyl 2-{2-methyl-4-[({4-methyl-2-[3,4-dichlorophenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetate
2-{2-methyl-4-[({4-methyl-2-[3,5-bis(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetic acid
ethyl 2-{2-methyl-4-[({4-methyl-2-[3,5-bis(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetate
ethyl 2-{2-methyl-4-[({4-methyl-2-[3-fluoro-4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetate
5-[4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-oxazol-5-yl}methoxy)phenoxymethyl]-2H-tetrazole
5-[4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-oxazol-5-yl}methoxy)benzyl]-2H-tetrazole
5-[4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}sulfanyl)benzyl]-2H-tetrazole
5-[4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-oxazol-5-yl}sulfanyl)benzyl]-2H-tetrazole
5-[2-methyl-4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methoxy)benzyl]-2H-tetrazole
5-[2-methyl-4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}sulfanyl)benzyl]-2H-tetrazole
More preferred compounds of the invention are:
2-{2-methyl-4-[({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetic acid
2-{2-methyl-4-[({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-oxazol-5-yl}methyl)sulfanyl]phenoxy}acetic acid
methyl 2-{4-[({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetate
2-{4-[({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetic acid
(E)-3-[2-methyl-4-({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methoxy)phenyl]-2-propenoic acid
2-{3-chloro-4-[({4-methyl-2-[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenyl}acetic acid
2-{2-methyl-4-[({4-methyl-2-[3-fluoro-4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetic acid
A particularly preferred compound of the invention is:
2-{2-methyl-4[({4-methyl-2[4-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methyl)sulfanyl]phenoxy}acetic acid.
All the preferred and most preferred compounds listed above are selective hPPARxcex4 agonists.
It will also be appreciated by those skilled in the art that the compounds of the present invention may also be utilized in the form of a pharmaceutically acceptable salt or solvate thereof. The physiologically acceptable salts of the compounds of formula (I) include conventional salts formed from pharmaceutically acceptable inorganic or organic acids or bases as well as quatemary ammonium acid addition salts. More specific examples of suitable acid salts include hydrochloric, hydrobromic, sulfuric, phosphoric, nitric, perchloric, fumaric, acetic, propionic, succinic, glycolic, formic, lactic, maleic, tartaric, citric, palmoic, malonic, hydroxymaleic, phenylacetic, glutamic, benzoic, salicylic, fumaric, toluenesulfonic, methanesulfonic, naphthalene-2-sulfonic, benzenesulfonic hydroxynaphthoic, hydroiodic, malic, steroic, tannic and the like. Other acids such as oxalic, while not in themselves pharmaceutically acceptable, may be useful in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable salts. More specific examples of suitable basic salts include sodium, lithium, potassium, magnesium, aluminium, calcium, zinc, N,Nxe2x80x2-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucamine and procaine salts. Those skilled in the art of organic chemistry will appreciate that many organic compounds can form complexes with solvents in which they are reacted or from which they are precipitated or crystallized. These complexes are known as xe2x80x9csolvatesxe2x80x9d. For example, a complex with water is known as a xe2x80x9chydratexe2x80x9d. Solvates of the compound of formula (I) are within the scope of the invention. References hereinafter to a compound according to the invention include both compounds of formula (I) and their pharmaceutically acceptable salts and solvates.
It will be appreciated by those skilled in the art that reference herein to treatment extends to prophylaxis as well as the treatment of established diseases or symptoms. Moreover, it will be appreciated that the amount of a compound of the invention required for use in treatment will vary with the nature of the condition being treated and the age and the condition of the patient and will be ultimately at the discretion of the attendant physician or veterinarian. In general, however, doses employed for adult human treatment will typically be in the range of 0.02-5000 mg per day, preferably 1-1500 mg per day. The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example as two, three, four or more sub-doses per day.
While it is possible that compounds of the present invention may be therapeutically administered as the raw chemical, it is preferable to present the active ingredient as a pharmaceutical formulation. Accordingly, the present invention further provides for a pharmaceutical formulation comprising a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof together with one or more pharmaceutically acceptable carriers therefor and, optionally, other therapeutic and/or prophylactic ingredients.
Formulations of the present invention include those especially formulated for oral, buccal, parenteral, transdermal, inhalation, intranasal, transmucosal, implant, or rectal administration, however, oral administration is preferred. For buccal administration, the formulation may take the form of tablets or lozenges formulated in conventional manner. Tablets and capsules for oral administration may contain conventional excipients such as binding agents, (for example, syrup, acacia, gelatin, sorbitol, tragacanth, mucilage of starch or polyvinylpyrrolidone), fillers (for example, lactose, sugar, microcrystalline cellulose, maize-starch, calcium phosphate or sorbitol), lubricants (for example, magnesium stearate, stearic acid, talc, polyethylene glycol or silica), disintegrants (for example, potato starch or sodium starch glycollate) or wetting agents, such as sodium lauryl sulfate. The tablets may be coated according to methods well-known in the art.
Alternatively, the compounds of the present invention may be incorporated into oral liquid preparations such as aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, for example. Moreover, formulations containing these compounds may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations may contain conventional additives such as suspending agents such as sorbitol syrup, methyl cellulose, glucose/sugar syrup, gelatin, hydroxyethylcellulose, carboxymethyl cellulose, aluminum stearate gel or hydrogenated edible fats; emulsifying agents such as lecithin, sorbitan mono-oleate or acacia; non-aqueous vehicles (which may include edible oils) such as almond oil, fractionated coconut oil, oily esters, propylene glycol or ethyl alcohol; and preservatives such as methyl or propyl p-hydroxybenzoates or sorbic acid. Such preparations may also be formulated as suppositories, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.
Additionally, formulations of the present invention may be formulated for parenteral administration by injection or continuous infusion. Formulations for injection may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle (e.g., sterile, pyrogen-free water) before use.
The formulations according to the invention may also be formulated as a depot preparation. Such long acting formulations may be administered by implantation (for example, subcutaneously or intramuscularly) or by intramuscular injection. Accordingly, the compounds of the invention may be formulated with suitable polymeric or hydrophobic materials (as an emulsion in an acceptable oil, for example), ion exchange resins or as sparingly soluble derivatives as a sparingly soluble salt, for example.
The formulations according to the invention may contain between 0.1-99% of the active ingredient, conveniently from 30-95% for tablets and capsules and 3-50% for liquid preparations.
The compound of formula (I) for use in the instant invention may be used in combination with other therapeutic agents for example, statins and/or other lipid lowering drugs for example MTP inhibitors and LDLR upregulators. The compounds of the invention may also be used in combination with antidiabetic agents, e.g. metformin, sulfonylureas, or PPAR gamma, PPAR alpha and PPAR alpha/gamma agonists (for example thiazolidinediones such as e.g. Pioglitazone and Rosiglitazone). The compounds may also be used in combination with antihypertensive agents such as angiotensin antagonists eg telmisartan, calcium channel antagonists eg lacidipine and ACE inhibitors eg enalapril. The invention thus provides in a further aspect the use of a combination comprising a compound of formula (I) with a further therapeutic agent in the treatment of a hPPAR delta mediated disease.
When the compounds of formula (I) are used in combination with other therapeutic agents, the compounds may be administered either sequentially or simultaneously by any convenient route.
The combinations referred to above may conveniently be presented for use in the form of a pharmaceutical formulation and thus pharmaceutical formulations comprising a combination as defined above optimally together with a pharmaceutically acceptable carrier or excipient comprise a further aspect of the invention. The individual components of such combinations may be administered either sequentially or simultaneously in separate or combined pharmaceutical formulations.
When combined in the same formulation it will be appreciated that the two compounds must be stable and compatible with each other and the other components of the formulation and may be formulated for administration. When formulated separately they may be provided in any convenient formulation, conveniently in such a manner as are known for such compounds in the art.
When a compound of formula (I) is used in combination with a second therapeutic agent active against the same disease, the dose of each compound may differ from that when the compound is used alone. Appropriate doses will be readily appreciated by those skilled in the art.
Compounds of this invention may be conveniently prepared by a general process wherein a moiety like A is coupled to an alcohol (B) using the Mitsunobu protocol (O. Mitsunobu, 1981 Synthesis, p 1) or by alkylation of A using a suitable non nucleophilic base such as K2CO3, Cs2CO3 or NaH, with an alkyl halide (C). Note that this synthesis is preferably carried out with the acid group protected by R. Preferably, R is 1-6 alkyl which can be hydrolyzed off to give an acid of Formula (I), or if readily hydrolyzable, the resulting ester can be administered. 
For example, when n is 1, Y is S, Z is N, and R3 is para-CF3: 
Some of the intermediates of type A are commercially available while others can be synthesized as outlined below. The synthesis of intermediates of type B is also illustrated below.
Furthermore, the tetrazole derivatives may be conveniently prepared by a general process wherein a moiety like D is coupled to an alcohol (B) using the Mitsunobu protocol (O. Mitsunobu, 1981 Synthesis, p 1), by alkylation of D using a suitable non nucleophilic base such as K2CO3, Cs2CO3 or NaH, with an alkyl halide (C) or by coupling of a moiety like E with an alkyl halide (C) using a suitable non nucleophilic base such as NaOH. 
For example, when n is 1, Y is S, Z is N, and R3 is para-CF3: 
The invention is further illustrated by the following Intermediates and Examples which should not be construed as constituting a limitation thereto.

To a well stirred solution of LiAlH4 (1.52 g, 40 mmol) in dry THF (50 mL) at 0xc2x0 C., was slowly added a solution of ethyl 4-methyl-2-[4-(trifluoromethyl)phenyl]-thiazole-5-carboxylate (12.6 g, 40 mmol) in dry THF (50 mL). The mixture was stirred at room temperature for 2 hs. The reaction was quenched by slow addition at 0xc2x0 C. of water (2 mL), 5N NaOH (2 mL) and water (6 mL). The precipitate was filtered, washed with EtOAc, MeOH, CH2Cl2 and THF. After evaporation, a yellow solid was obtained, that was crystallized from MeOH-water to afford intermediate 1 depicted above (9.90 g, 36 mmol, 90%) as a yellow solid mp 120-122xc2x0 C. 
To a cold (0xc2x0 C.) stirred solution of intermediate 1 (8.2 g, 30 mmol) and Et3N (6.07 g, 8.36 mL, 60 mmol), in dry CH2Cl2 (120 mL) was slowly added MeSO2Cl (5.49 g, 3.71 mL, 48 mmol). After 2 hs at 0xc2x0 C. more Et3N (6 mmol) and MeSO2Cl (4.8 mmol) were added. After 2 more h a tic (hexane:EtOAc, 1:1) showed complete reaction. The reaction mixture was diluted with CH2Cl2 (120 mL) and washed with NaHCO3 (sat.) (2xc3x97240 mL) and water (2xc3x97240 mL), dried, filtered and evaporated to afford intermediate 2 (8.0 g, 27 mmol, 90%) as a yellow solid. 
A neat mixture of methyl 2-chloroacetoacetate (9.88 g, 8.0 mL, 65.6 mmol) and 4-(trifluoromethyl)benzamide acid (5.67 g, 30 mmol), was heated in an oil bath at 120xc2x0 C. for 48 h. The dark mixture was cooled down to room temperature and diluted with EtOAc (100 mL) and succesively washed with: NaHCO3 (sat.) (3xc3x97100 mL) and water (3xc3x97100 mL), dried, filtered and evaporated to a syrup. The syrup was dissolved in acetone and precipitated with hexane. The solids (unreacted 4-(trifluoromethyl)benzamide acid were filtered and washed with more hexane. The solution was evaporated under vacuum at 60xc2x0 C. to eliminate traces of methyl 2-chloroacetoacetate. The resulting mixture was purified by flash column chromatography (hexane:EtOAc, 95:5), to afford intermediate 3 (2.2 g, 7.7 mmol, 25%) as a white solid. 
To a well stirred solution of LiAlH4 (213 mg, 5.6 mmol) in dry THF (7.0 mL) at 0xc2x0 C., was slowly added a solution of intermediate 3 (1.6 g, 5.6 mmol) in dry THF (7.0 mL). The mixture was stirred at room temperature for 2 h. The reaction was quenched by slow addition at 0xc2x0 C. of water (0.3 mL), 5N NaOH (0.3 mL) and water (0.9 mL). The precipitate was filtered, washed with EtOAc, i MeOH, CH2Cl2 and THF. After evaporation, intermediate 4 (1.1 g, 4.3 mmol, 77%) was obtained as a yellow solid. 
To a cold (0xc2x0 C.) stirred solution of intermediate 4 (2.57 g, 10 mmol) and Et3N (2.02 g, 2.78 mL, 20 mmol), in dry CH2Cl2 (40 mL) was slowly added MeSO2Cl (1.83 g, 1.24 mL, 16 mmol). After 2 h at 0xc2x0 C. more Et3N (4 mmol) and MeSO2Cl (3.2 mmol) were added. After 2 more h a tic (hexane:EtOAc, 1:1), showed complete reaction. The reaction mixture was diluted with CH2Cl2 (40 mL) and washed with NaHCO3 (sat.) (2xc3x9780 mL) and water (2xc3x9780 mL), dried, filtered and evaporated to afford intermediate 5 (2.8 g, 10 mmol, 100%) as a yellow solid. 
A solution of 3-(trifluoromethyl)benzoic acid (570 mg, 3.0 mmol) in dry DMF (10 mL) was heated at 55xc2x0 C. Solid K2CO3 (220 mg, 1.53 mmol) was added, followed by methyl 2-chloroacetoacetate (452 mg, 3.0 mmol). The suspension was stirred for 1.5 h at 55xc2x0 C. The reaction was then partitioned between water (40 mL) and ether (50 mL). The organic layer was further washed with brine (2xc3x9740 mL), dried, filtered and evaporated to a yellow oil. A solution of this oil in AcOH (10 mL), was added to a suspension of NH4OAc (0.64 g, 8.3 mmol) in dry toluene (10 mL). The reaction was then refluxed overnight. It was poured into ice/water (60 mL) and extracted with ether (4xc3x9730 mL). The organic layer was washed with brine (2xc3x9760 mL), dried, filtered and evaporated to give crude material that was purified by flash column chromatography (CH2Cl2) to afford intermediate 6 (320 mg, 1.12 mmol, 37%) as a white solid. 
To a well stirred solution of LIAlH4 (38 mg, 1.0 mmol) in dry THF (1.5 mL) at 0xc2x0 C., was slowly added a solution of intermediate 6 (285 mg, 1.0 mmol) in dry. THF (1.5 mL). The mixture was stirred at room temperature for 2 h. The reaction was quenched by slow addition at 0xc2x0 C. of water (100 xcexcL), 5N NaOH (100 xcexcL) and water (300 xcexcL). The precipitate was filtered, washed with EtOAc, MeOH, CHl2C2 and THF to afford intermediate 7 (210 mg, 0.82 mmol, 82%) as a white solid. 
A solution of benzoic acid (1.22 g, 10.0 mmol) in dry DMF (20 mL) was heated at 55xc2x0 C. Solid K2CO3 (691 mg, 5.0 mmol) was added, followed by methyl 2-chloroacetoacetate (1.50 g, 10.0 mmol). The suspension was stirred for 1.5 h at 55xc2x0 C. The reaction was then partitioned between water (150 mL) and ether (150 mL). The organic layer was further washed with brine (2xc3x97150 mL), dried, filtered and evaporated to a yellow oil. A solution of this oil in AcOH (20 mL), was added to a suspension of NH4OAc (2.13 g, 28 mmol) in dry toluene (20 mL). The reaction was then refluxed overnight. It was poured into ice/water (200 mL) and extracted with ether (4xc3x97100 mL). The organic layer was washed with brine (2xc3x97200 mL), dried, filtered and evaporated to give crude material that was purified by flash column chromatography (hexane:EtOAc, 4:1) to afford intermediate 8 (720mg, 3.13 mmol, 32%) as a white solid. 
To a well stirred solution of LiAlH4 (76 mg, 2.0 mmol) in dry THF (2.5 mL) at 0xc2x0 C., was slowly added a solution of intermediate 8 (434 mg, 2.0 mmol) in dry THF (2.5 mL). The mixture was stirred at room temperature for 2 h. The reaction was quenched by slow addition at 0xc2x0 C. of water (100 xcexcL), 5N NaOH (100 xcexcL) and water (300 xcexcL). The precipitate was filtered, washed with EtOAc, 10 MeOH, CH2Cl2 and THF to afford intermediate 9 (349 mg, 0.92 mmol, 46%). Which was used without further purification. 
A solution of ethyl-3-bromo-4-oxopentanoate (670 mg, 3.0 mmol) and 4-(trifluoromethyl)thiobenzamide (677 mg, 3.3 mmol) in EtOH (5 mL) was refluxed overnight. After cooling to room temperature the solution was diluted with AcOEt. After adding hexane a precipitate appeared. It was filtered and washed with hexane to afford intermediate 10 (300 mg, 0.91 mmol) as a white solid. The mother liquors were evaporated to a syrup that was purified by flash column chromatography (hexane:EtOAc, 9:1) to afford additional intermediate 10 (300 mg, 0.91 mmol). Total yield was 60%. 
To a well stirred solution of LiAlH4 (31 mg, 0.8 mmol) in dry THF (1.0 mL) at 0xc2x0 C., was slowly added a solution of intermediate 10 (264 mg, 0.8 mmol) in dry THF (1.5 mL) and dry CH2Cl2 (1.5 mL). The mixture was stirred at room temperature for 2 h. The reaction was quenched by slow addition at 0xc2x0 C. of water (50.0 xcexcL), 5N NaOH (50.0 xcexcL) and water (150 xcexcL). The precipitate was filtered, washed with EtOAc, MeOH, CH2Cl2 and THF. After evaporation intermediate 11 (133 mg, 0.46 mmol, 57%) was obtained as a yellow solid. 
A solution of methyl-4-bromo-3-oxopentanoate (890 mg, 4.0 mmol) and and 4-(trifluoromethyl)thiobenzamide (820 mg, 4.0 mmol) in EtOH (10 mL) was refluxed overnight. After cooling to room temperature the solution was diluted with AcOEt and successively washed with (sat.) NaHCO3 (3xc3x9750 mL) and brine (2xc3x9750 mL), dried, filtered, and evaporated to dryness. A yellow solid was obtained, that was purified by flash column chromatography (hexane:EtOAc, 1:1) to afford the title compound (1.32 g, 4.0 mmol, 100%) as a white solid. 
To a well stirred solution of LiAlH4 (76 mg, 2.0 mmol) in dry THF (2.5 mL) at 0xc2x0 C., was slowly added a solution of Intermediate 12 (659 mg, 2.0 mmol) in dry THF (2.5 mL). The mixture was stirred at room temperature for 2 h. The reaction was quenched by slow addition at 0xc2x0 C. of water (100 xcexcL), SN NaOH (100 xcexcL) and water (300 xcexcL). The precipitate was filtered, washed with EtOAc, MeOH, CH2Cl2 and THF. After evaporation compound the title compound was obtained as a yellow solid (472 mg, 1.64 mmol, 82%). 
Methyl acrylate and 4-bromo-3-methylphenol were coupled using Heck conditions as described in the general procedure 4. The crude material was crystallized from acetone:hexane to afford the title compound (40%) as an amorphous solid. 
A solution of intermediate 14 (1.92 g, 10 mmol) in EtOAc (50 mL) was A hydrogenated at 50-60 psi at room temperature, in the presence of Pd/C 10% (500 mg). After 15 min., the mixture was filtered through celite, washed with additional EtOAc and evaporated to afford the title compound (1.94 g, 10 mmol, 100%) as a colorless syrup. 
N-Methylanisidine (2.0 g, 15 mmol), methyl bromoacetate (2.25 g, 15 mmol), DMAP (0.04 g, 2% by wt), and Net3 (2.25 g, 15 mmol) in EtOH (50 mL) was refluxed for 1 h. The solvents were evaporated. The remaining residue was chromatographed on a silica gel column with 10% EtOAc in hexanes to afford the title compound (70%) as a yellow oil: NMR (DMSO-d6) xcex4 2.97 (s, 3H), 3.66 (s, 3H), 3.72 (s, 3H), 4.18 (s, 2H), 6.68 (d, 2H), 6.85 (d, 2H).
MS m/z 210 (M+1)xe2x88x92. 
Intermediate 16 (1.9 g, 9.0 mmol) in CH2Cl2 (10 mL) was added to 1M BBr3 in CH2Cl2 (28 mL) slowly at 0xc2x0 C. The resulting solution was stirred at low temperature for 2 h, and poured onto ice-water. The mixture was extracted with CH2CH2 (2xc3x9750 mL), dried, and evaporated. A solution of this residue and acetyl chloride (1.4 g, 18 mmol) in MeOH was refluxed for 18 h. The solvents were evaporated. The residue was chromatographed over silica gel to afford the title compound (45%) as a yellow oil: NMR (MeOH-d4) xcex4 3.39 (s, 3H), 3.72 (s, 3H), 4.51 (s, 2H), 6.87 (d, 2H), 7.48 (d, 2H).
MS m/z 196(M+1)xe2x88x92. 
Methyl 3-Chloro-4-hydroxyphenylacetate was treated with dimethyl thiocarbamoyl chloride as described in general procedure 5 to afford, after column chromatography (hexane:EtOAc, 4:1), a brown oil (95%). The residue was refluxed in tetradecane as to afford after column chromatography (hexane:EtOAc, 4:1) the title compound (77%) as a yellow oil. 
Methyl 2-(4-hydroxyphenoxy)acetate was treated with dimethyl thiocarbamoyl chloride as described in general procedure 5 to afford, after column chromatography (hexane:EtOAc, 4:1), (84%) as a yellow oil. The oil was refluxed in tetradecane as to afford after column chromatography (hexane:EtOAc, 4:1) the title compound (53%) as a yellow oil. 
A mixture of methyl bromoacetate (3.80 g, 2.35 mL, 25.0 mmol), 4-hydroxy-3-methylacetophenone (4.13 g, 27.5 mmol), and Cs2CO3 (17.9 g, 55 mmol) in dry acetonitrile (125 mL) was stirred overnight at r.t. The mixture was filtered, washed with acetonitrile, and the solvent evaporated. The remaining syrup was redissolved in EtOAc (400 mL), washed with 1N NaOH (3xc3x97400 mL) and water (2xc3x97400 mL), dried, filtered, and evaporated to afford the pure title compound (5.50 g, 24.7 mmol, 99%) as a white solid. 
A solution of Intermediate 20 (5.33 g, 24 mmol), mCPBA (7.25 g, 42 mmol) and p-TsOH (480 mg) in dry dichloromethane (120 mL) was refluxed for 48 h. The reaction mixture was diluted with dichloromethane (120 mL), and successively washed with: aq. Kl (2xc3x97200 mL), NaHSO3 (2xc3x97200 mL), dried, filtered and evaporated to afford the title compound (5.0 g, 21 mmol, 87%) as a syrup. 
A solution of intermediate 21 (4.76 g, 20 mmol) in dry methanol (180 mL) was treated with a 0.5 N solution of NaOCH3 in MeOH (40 mL, 20 mmol). After 1 h at r.t., the solution was neutralized with 1N HCl (20 mL). The solvent was evaporated, and the residue partitioned between dichloromethane (300 mL) and water (300 mL). The organic solution was separated, washed with water (300 mL), dried, filtered, and evaporated to afford the title compound (3.3 g, 16.8 mmol, 84%) as a brown solid. 
Methyl 4-hydroxyphenylacetate was treated with dimethyl thiocarbamoyl chloride as described in general procedure 5 to afford, after column chromatography (hexane:EtOAc, 4:1) (90%) a yellow solid. The solid was refluxed in tetradecane to afford after column chromatography (hexane:EtOAc, 4:1) the title compound (74%) as a brown oil. 
Methyl 3-methoxy-4-hydroxyphenylacetate was treated with dimethyl thiocarbamoyl chloride as described in general procedure 5 to afford, after column chromatography (hexane:EtOAc, 4:1), a brown oil (95%). The oil was refluxed in tetradecane to afford after column chromatography (hexane:EtOAc, 4:1) compound the title compound (17%) as a yellow oil. 
Intermediate 22 was treated with dimethyl thiocarbamoyl chloride as described in general procedure 5 to afford a dark oil (100%). The dark oil was refluxed in tetradecane to afford after column chromatography (hexane:EtOAc, 2:1) compound a brown solid (47%). The brown solid was treated with NaOMe/HOMe to afford, after column chromatography (hexane:EtOAc, 4:1), compound the title compound (34%) as a colorless syrup.
To a solution of P4S10 (0.2 mmol) in toluene (100 mL) was added NaHCO3 (2 mmol) and the mixture heated to reflux for ca. 30 min. The substituted benzamide (1 mmol) was added and the reaction stirred at 90xc2x0 C. for 1 h. The reaction was then evaporated to dryness, treated with brine (100 mL) and extracted with CH2Cl2 (2xc3x9750 mL). The organic phase dried, filtered, and evaporated to afford the final product. 
The title compound was prepared as described in general procedure A to afford an orange solid (88%).
MS m/z 217 (M+1). 
The title compound was prepared as described in general procedure A to afford an orange solid (99%).
MS m/z 171. 
The title compound was prepared as described in general procedure A to afford an orange solid (58%).
MS m/z 155. 
The title compound was prepared as described in general procedure A to afford a yellow solid (87%).
MS m/z 207 (M+1). 
The title compound was prepared as described in general procedure A to afford a brownish orange solid (78%).
MS m/z 173. 
The title compound was prepared as described in general procedure A to afford a yellow semi-solid (55%).
MS m/z 273. 
The title compound was prepared as described in general procedure A to afford a yellow solid (50%).
MS m/z 223.
To a solution of the substituted thiobenzamide (1 mmol) in EtOH (100 mL) was added ethyl 2-chloroacetoacetate (1.1 mmol) and the mixture heated to reflux overnight. The reaction is cooled to room temperature and the solvent evaporated. The solid is crystallized from Et2O or hexane to afford the final product. 
Intermediate 26 was reacted as described in general procedure B to afford the title compound as an off-white solid (41%).
MS m/z 327 (M+1). 
Intermediate 27 was reacted as described in general procedure B to afford the title compound as an off-white solid (29%).
MS m/z 281. 
Intermediate 28 was reacted as described in general procedure B to afford the title compound as an off-white solid (25%).
1H-NMR (CDCl3) xcex4 1.35 (t, 3H), 2.75 (s, 3H), 4.35 (q, 2H), 7.15 (t, 2H), 7.95 (dd, 2H). 
Intermediate 29 was reacted as described in general procedure B to afford the title compound as an off-white solid (46%).
MS m/z 315. 
Intermediate 30 was reacted as described in general procedure B to afford the title compound as an off-white solid (41%).
1H-NMR (CDCl3) xcex4 1.35 (t, 3H), 2.75 (s, 3H), 4.35 (q, 2H), 7.25 (dd, 1H), 7.65 (m, 1H), 7.75 (ddd, 1H). 
Intermediate 31 was reacted as described in general procedure B to afford the title compound as an off-white solid (58%).
MS m/z 383. 
Intermediate 32 was reacted as described in general procedure B to afford the title compound as an off-white solid (56%).
MS m/z 333.
To a solution of LiAlH4 (2 mmol) in THF (100 mL) at 0xc2x0 C. was added the 2-substituted phenyl-4-methyl-1,3-thiazole-5-carboxylic acid ethyl ester. The reaction is stirred while it is allowed to warm to rt. After all the starting material has disappeared, the reaction is cautiously treated with water (5 mL) followed by 1N NaOH (10 mL). The mixture was filtered through celite. The filtrate was extracted with CH2Cl2 (3xc3x9750 mL). The organic phase was dried, filtered and 15 evaporated to afford the final product. 
Intermediate 33 was reacted as described in general procedure C to afford the title compound as an off-white solid (75%).
MS m/z 285 (M+1). 
Intermediate 34 was reacted as described in general procedure C to afford the title compound as an off-white solid (87%).
MS m/z 239. 
Intermediate 35 was reacted as described in general procedure C to afford the title compound as an off-white solid (89%).
1H-NMR (CDCl3) xcex4 1.7 (bs, 1H), 2.35 (s, 3H), 4.75 (s, 2H), 7.05 (t, 2H), 7.80 (dd, 2H). 
Intermediate 36 was reacted as described in general procedure C to afford the title compound as an off-white solid (56%).
MS m/z 275 (M+1). 
Intermediate 37 was reacted as described in general procedure C to afford the title compound as an off-white solid (52%).
MS m/z 241. 
Intermediate 38 was reacted as described in general procedure C to afford the title compound as an off-white solid (27%).
MS m/z 341. 
Intermediate 39 was reacted as described in general procedure C to afford the title compound as an off-white solid (63%).
MS m/z 291.
To a solution of the 2-substituted phenyl-5-hydroxymethyl-4-methyl-1,3-thiazole (1 mmol) and Et3N (2 mmol) in CH2Cl2 (100 mL) at 0xc2x0 C. was added dropwise methanesulfonyl chloride (1.6 mmol). After 2-4 h the reaction was complete. CH2Cl2 (50 mL) is added and the organic phase washed with a saturated. NaHCO3 solution (2xc3x9750 mL), water (2xc3x9750 mL), dried, filtered and then evaporated to afford the final product. 
Intermediate 40 was reacted as described in general procedure D to afford the title compound as an white solid (40%).
MS m/z 303. 
Intermediate 41 was reacted as described in general procedure D to afford the title compound as an white solid (80%).
MS m/z 259 (M+1). 
Intermediate 42 was reacted as described in general procedure D to afford the title compound as a pale yellow solid (100%).
MS m/z 241. 
Intermediate 43 was reacted as described in general procedure D to afford the title compound as a pale yellow solid (74%).
1H-NMR (CDCl3) xcex4 2.40 (s, 3H), 4.70 (s, 2H), 7.40 (dd, 1H), 7.60 (dd, 1H), 7.90 (d, 1H). 
Intermediate 44 was reacted as described in general procedure D to afford the title compound as a pale yellow solid (83%).
1H-NMR (CDCl3) xcex4 2.30 (s, 3H), 4.60 (s, 2H), 7.00 (dd, 1H), 7.40 (m, 1H), 7.50 (m, 1H). 
Intermediate 45 was reacted as described in general procedure D to afford the title compound as a pale yellow solid (100%).
1H-NMR (CDCl3) xcex4 2.40 (s, 3H), 4.70 (s, 2H), 7.80 (s, 1H), 8.30 (s, 2H). 
Intermediate 46 was reacted as described in general procedure D to afford the title compound as a pale yellow solid (100%).
1H-NMR (CDCl3) xcex4 2.40 (s, 3H), 4.70 (s, 2H), 7.55-7.75 (m, 3H). 
Chlorosulfonic acid (15 mL) was cooled to 0xc2x0 C. then 10.0 g (0.05M) of ethyl (2-methylphenoxyacetate was added over 10 m. The reaction mixture was stirred at 0-5xc2x0 C. for 30 m, the bath was removed and stirring continued for 2 h. The reaction mixture was poured into ice, forming a white solid which was washed with ice water and dried under high vacuum affording the title compound (12.846 g, 86%). 
2-Fluoro-4-(trifluoromethyl)benzenecarbothioamide
To a solution of 2-fluoro-4-(trifluoromethyl)benzonitrile (5.2 g, 27.5 mmol) in 50 ml methanol was added 10 ml of water and NaSHxc3x97H2O (7.71 g, 137.5 mmol). After heating at 50xc2x0 C. for 12 hours, the solvent was removed in vacuo and the residue treated with water (200 ml) and extracted with ethyl acetate (2xc3x97150 ml).
The organic layers were dried (MgSO4) and the solvent evaporated to give crude residue which was purified by Biotage FlashElute with a 40M silica cartridge and eluting with hexanes/ethyl acetate (4:1)
To yield 3.27 g (53%) of 2-fluoro-4-(trifluoromethyl)benzenecarbothioamide, intermediate 55 as a yellow solid. MS m/z 223 (M+1). 
Intermediate 55 was reacted as described in general procedure B to afford the title compound as a light yellow solid (71%)
MS m/z 333 (M+1). 
Intermediate 56 was reacted as described in general procedure C to afford the title compound as a light yellow solid (83%)
MS m/z 291 (M+1). 
Intermediate 57 was reacted as described in general procedure D to afford the title compound as a light yellow solid (100%)
Rf of starting alcohol in 3:1 hexanes/ethyl acetate is 0.25.
Rf of chloride in 3:1 hexanes/ethyl acetate is 0.75. 
4-hydroxybenzyl cyanide was treated with dimethyl thiocarbamoyl chloride as described in general procedure 5 to afford, after column chromatography (DCM), a yellow solid (78%). The solid was refluxed in tetradecane as to afford after column chromatography (DCM:MeOH) the title compound (40%) as an off-white solid. 
4-hydroxy-2-methylbenzyl cyanide was treated with dimethyl thiocarbamoyl chloride as described in general procedure 5 to afford, after column chromatography (DCM), a yellow solid (48%). The solid was refluxed in tetradecane as to afford after column chromatography (DCM) the title compound (60%) as an off-white solid. 
A solution of intermediate 59 (1 g, 4.5 mmol) and NaOH (0.2 g, 5 mmol) in dry MeOH (20 mL) was heated at 70xc2x0 C. for 5 h. Then, intermediate 5 (1.25 g, 4.5 mmol) was added and the reaction was stirred at 70xc2x0 C. for one more hour and 18 hours at room temperature. After evaporation of the solvent, the residue was purified by flash column chromatography (DCM) to afford the title compound (71%) as a yellow oil. 
Intermediate 4 and 4-hydroxyphenylacetonitrile were coupled as described in the general procedure 1 to afford the title compound (56%) as a yellow oil. 
Intermediate 4 and 4-hydroxybenzyl cyanide were coupled as described in the general procedure 1 to afford the title compound (39%) as a pale yellow solid. 
A solution of intermediate 59 (1 g, 4.5 mmol) and NaOH (0.2 g, 5 mmol) in dry MeOH (20 mL) was heated at 70xc2x0 C. for 5 h. Then, intermediate 2 (1.65 g, 4.5 mmol) was added and the reaction was stirred at 70xc2x0 C. for one more hour and 18 hours at room temperature. After evaporation of the solvent, the residue was purified by flash column chromatography (DCM) to afford the title compound (75%) as a yellow oil. 
Intermediate 2 and 4-hydroxy-2-methylbenzyl cyanide were coupled as described in the general procedure 1 to afford the title compound (26%) as a white yellow solid. 
A solution of intermediate 60 (1.67 g, 7.1 mmol) and NaOH (0.32 g, 7.8 mmol) in dry MeOH (20 mL) was heated at 85xc2x0 C. for 5 h. Then, intermediate 2 (1.65 g, 4.5 mmol) was added and the reaction was stirred at 85xc2x0 C. for one more hour and 18 hours at room temperature. After evaporation of the solvent, the residue was purified by flash column chromatography (DCM) to afford the title compound (20%) as a yellow oil.